Eastern North America is considered the center of diversity for many Vitis spp. and for the grape powdery mildew pathogen, Erysiphe necator. However, little is known about populations of E. necator from wild Vitis spp. We determined the phenotypic variation in pathogenicity and aggressiveness of E. necator among isolates from wild and domesticated Vitis spp. from diverse geographic regions in the eastern United States. To test pathogenicity, we inoculated 38 E. necator isolates on three wild Vitis spp., two commercially grown hybrids and the European wine grape, Vitis vinifera. V. rotundifolia (muscadine grape) was the only host species on which complete host specialization was evident; it was only susceptible to isolates collected from V. rotundifolia. All isolates, regardless of source host, were pathogenic on the other Vitis spp. We found no differences in components of aggressiveness latent period and lesion size among isolates from different source hosts when inoculated on V. vinifera, which is highly susceptible to powdery mildew. However significant variation was evident among isolates on the more resistant V. labruscana 'Niagara'. Isolates from the wild species V. aestivalis were the most aggressive, whereas isolates from V. vinifera were not more aggressive than isolates from other source hosts. Greater aggressiveness was also detected among isolates from the southeastern United States compared with isolates from the northeastern United States.